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tak  (MedChemExpress)


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    MedChemExpress tak
    ( A to C ) HIEs were pretreated for 3 hours with the indicated concentrations <t>of</t> <t>TAK-779</t> and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the absence or presence of the same concentrations of TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from two independent experiments (J2 and J8FUT2-KI HIEs) and three independent experiments (J4FUT2-KI HIEs) with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Significance was determined using two-way ANOVA with Tukey’s post hoc test comparing TAK-779 concentrations and individual experimental results at 48 hours postinfection. P values are shown for individual TAK-779 concentrations versus no TAK-779 treatment control (* P value <0.05, ** P < 0.01, *** P < 0.001, and ****, 0.0001). Significant differences between experiments were observed for the studies conducted in J4FUT2-KI and J8FUT2-KI HIEs ( P < 0.0001 for each).
    Tak, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids"

    Article Title: Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids

    Journal: Science Advances

    doi: 10.1126/sciadv.aeb0455

    ( A to C ) HIEs were pretreated for 3 hours with the indicated concentrations of TAK-779 and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the absence or presence of the same concentrations of TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from two independent experiments (J2 and J8FUT2-KI HIEs) and three independent experiments (J4FUT2-KI HIEs) with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Significance was determined using two-way ANOVA with Tukey’s post hoc test comparing TAK-779 concentrations and individual experimental results at 48 hours postinfection. P values are shown for individual TAK-779 concentrations versus no TAK-779 treatment control (* P value <0.05, ** P < 0.01, *** P < 0.001, and ****, 0.0001). Significant differences between experiments were observed for the studies conducted in J4FUT2-KI and J8FUT2-KI HIEs ( P < 0.0001 for each).
    Figure Legend Snippet: ( A to C ) HIEs were pretreated for 3 hours with the indicated concentrations of TAK-779 and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the absence or presence of the same concentrations of TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from two independent experiments (J2 and J8FUT2-KI HIEs) and three independent experiments (J4FUT2-KI HIEs) with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Significance was determined using two-way ANOVA with Tukey’s post hoc test comparing TAK-779 concentrations and individual experimental results at 48 hours postinfection. P values are shown for individual TAK-779 concentrations versus no TAK-779 treatment control (* P value <0.05, ** P < 0.01, *** P < 0.001, and ****, 0.0001). Significant differences between experiments were observed for the studies conducted in J4FUT2-KI and J8FUT2-KI HIEs ( P < 0.0001 for each).

    Techniques Used: Infection, Cell Culture, Quantitative RT-PCR, Control

    Different conditions, pre- versus post-treatment (indicated) with pretreatment alone compared to post-treatment alone and 24 hours pretreatment, were compared. J4FUT2-KI HIEs were treated as indicated with 30 μM TAK-779. HIEs were then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the indicated conditions. After washing, the cells were cultured for 48 hours at 37 ° C in the presence of 30 μM TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from three independent experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Numbers above lines represent log 10 increase between no treatment versus indicated treatments. Significance was determined using two-way ANOVA with post hoc Tukey’s test comparing 48-hour replication, no TAK-779 to each condition of TAK-779 ( P value,**, < 0.001 and ****, 0.0001). hr, hours.
    Figure Legend Snippet: Different conditions, pre- versus post-treatment (indicated) with pretreatment alone compared to post-treatment alone and 24 hours pretreatment, were compared. J4FUT2-KI HIEs were treated as indicated with 30 μM TAK-779. HIEs were then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the indicated conditions. After washing, the cells were cultured for 48 hours at 37 ° C in the presence of 30 μM TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from three independent experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Numbers above lines represent log 10 increase between no treatment versus indicated treatments. Significance was determined using two-way ANOVA with post hoc Tukey’s test comparing 48-hour replication, no TAK-779 to each condition of TAK-779 ( P value,**, < 0.001 and ****, 0.0001). hr, hours.

    Techniques Used: Infection, Cell Culture, Quantitative RT-PCR

    ( A ) J4FUT2-KI HIEs were pretreated for 3 hours and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the presence of the indicated concentrations of TAK-779. After washing, the cells were cultured for the indicated times at 37°C in the presence of the indicated concentrations of TAK-779. Virus replication was evaluated by RT-qPCR at the indicated time points. ( B and C ) After 24 and 48 hours postinfection incubation, monolayers were fixed with methanol. GII.3-positive cells (green) were detected using guinea pig anti-HuNoV VLP Ab (green) and nuclei (blue) were detected by DAPI. (B) Representative images from each condition at 48 hours postinfection are shown. Scale bars denote 14 μm. (C) Numbers of HuNoV infected cells (Foci) per well from 24 and 48 hours postinfection condition were counted. Mean data compiled from two experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors using one-way ANOVA with post hoc Newman Keuls test comparing GEs per well for TAK-779 concentrations at 48- and 72-hour time points, ( P value, ****, 0.0001).
    Figure Legend Snippet: ( A ) J4FUT2-KI HIEs were pretreated for 3 hours and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the presence of the indicated concentrations of TAK-779. After washing, the cells were cultured for the indicated times at 37°C in the presence of the indicated concentrations of TAK-779. Virus replication was evaluated by RT-qPCR at the indicated time points. ( B and C ) After 24 and 48 hours postinfection incubation, monolayers were fixed with methanol. GII.3-positive cells (green) were detected using guinea pig anti-HuNoV VLP Ab (green) and nuclei (blue) were detected by DAPI. (B) Representative images from each condition at 48 hours postinfection are shown. Scale bars denote 14 μm. (C) Numbers of HuNoV infected cells (Foci) per well from 24 and 48 hours postinfection condition were counted. Mean data compiled from two experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors using one-way ANOVA with post hoc Newman Keuls test comparing GEs per well for TAK-779 concentrations at 48- and 72-hour time points, ( P value, ****, 0.0001).

    Techniques Used: Infection, Cell Culture, Virus, Quantitative RT-PCR, Incubation

    ( A ) J4FUT2-KI and ( B ) J8FUT2-KI HIEs were pretreated for 3 hours and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) with or without 30 μM TAK-779, labeled as P1. Following infection, both cells and supernatant were collected at 96 hours postinfection, and a portion was used to quantify viral RNA by RT-qPCR. The virus was subsequently used to infect a fresh batch of HIEs for the next passage, continuing for up to five passages. dpi, days postinfection; hpi, hours postinfection.
    Figure Legend Snippet: ( A ) J4FUT2-KI and ( B ) J8FUT2-KI HIEs were pretreated for 3 hours and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) with or without 30 μM TAK-779, labeled as P1. Following infection, both cells and supernatant were collected at 96 hours postinfection, and a portion was used to quantify viral RNA by RT-qPCR. The virus was subsequently used to infect a fresh batch of HIEs for the next passage, continuing for up to five passages. dpi, days postinfection; hpi, hours postinfection.

    Techniques Used: Infection, Labeling, Quantitative RT-PCR, Virus

    ( A ) J4FUT2-KI and J8FUT2-KI HIEs were pretreated and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the presence of 30 μM TAK-779 and indicated as P1. Postinfection, the cells and supernatant were harvested at 96 hours postinfection; after processing the supernatant, an aliquot was used to quantify the viral RNA by RT-qPCR. The virus was then used to infect a new batch of HIEs for the next passage, continuing up to 10 passages. ( B ) Immunofluorescence staining of HuNoV-infected J4FUT2-KI monolayers at P5 and P2. At 48 hours postinfection, cells were fixed with methanol and stained with guinea pig anti-HuNoV VLP antibody (green), rabbit anti-NTPase (red), and mouse anti-VPg (white). Nuclei were counterstained with DAPI (blue). Representative images are shown at high (10-μm scale bar) and low (20-μm scale bar) magnification. dpi, days postinfection; hpi, hours postinfection.
    Figure Legend Snippet: ( A ) J4FUT2-KI and J8FUT2-KI HIEs were pretreated and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the presence of 30 μM TAK-779 and indicated as P1. Postinfection, the cells and supernatant were harvested at 96 hours postinfection; after processing the supernatant, an aliquot was used to quantify the viral RNA by RT-qPCR. The virus was then used to infect a new batch of HIEs for the next passage, continuing up to 10 passages. ( B ) Immunofluorescence staining of HuNoV-infected J4FUT2-KI monolayers at P5 and P2. At 48 hours postinfection, cells were fixed with methanol and stained with guinea pig anti-HuNoV VLP antibody (green), rabbit anti-NTPase (red), and mouse anti-VPg (white). Nuclei were counterstained with DAPI (blue). Representative images are shown at high (10-μm scale bar) and low (20-μm scale bar) magnification. dpi, days postinfection; hpi, hours postinfection.

    Techniques Used: Infection, Quantitative RT-PCR, Virus, Immunofluorescence, Staining

    ( A ) J4FUT2-KI HIEs were pretreated for 3 hours with 30 μM TAK-779 and infected with GII.3 HuNoV derived from stool [2.9 × 10 5 genome equivalents (GEs) per well] or from stock 1 (2.58 × 10 5 GEs per well), stock 2 (2.93 × 10 5 GEs per well), stock 3 (1.02 × 10 5 GEs per well), or stock 4 (2.50 × 10 5 GEs per well). Infections were performed in the presence of 30 μM TAK-779 (left) or vehicle control (right). After washing, the cells were cultured for the indicated times at 37°C in the presence (left) or absence (right) of TAK-779. Virus replication was evaluated by RT-qPCR at the indicated time points. Viral replication was assessed by RT-qPCR. The characterization of each stock was performed once with three replicates. Error bars denote SD. ( B ) Representative images of stock-infected HIEs fixed with methanol at 48 hours postinfection. GII.3-positive cells were detected using guinea pig anti-HuNoV VLP Ab (green) and nuclei (blue) were detected by DAPI. Scale bars denote 14 μm. ( C ) Electron micrograph of HuNoV particles from the supernatant of HIEs 24 hours postinfection infected with stock virus. Scale bar, 50 nm; red line, ~40 nm, expected particle diameter.
    Figure Legend Snippet: ( A ) J4FUT2-KI HIEs were pretreated for 3 hours with 30 μM TAK-779 and infected with GII.3 HuNoV derived from stool [2.9 × 10 5 genome equivalents (GEs) per well] or from stock 1 (2.58 × 10 5 GEs per well), stock 2 (2.93 × 10 5 GEs per well), stock 3 (1.02 × 10 5 GEs per well), or stock 4 (2.50 × 10 5 GEs per well). Infections were performed in the presence of 30 μM TAK-779 (left) or vehicle control (right). After washing, the cells were cultured for the indicated times at 37°C in the presence (left) or absence (right) of TAK-779. Virus replication was evaluated by RT-qPCR at the indicated time points. Viral replication was assessed by RT-qPCR. The characterization of each stock was performed once with three replicates. Error bars denote SD. ( B ) Representative images of stock-infected HIEs fixed with methanol at 48 hours postinfection. GII.3-positive cells were detected using guinea pig anti-HuNoV VLP Ab (green) and nuclei (blue) were detected by DAPI. Scale bars denote 14 μm. ( C ) Electron micrograph of HuNoV particles from the supernatant of HIEs 24 hours postinfection infected with stock virus. Scale bar, 50 nm; red line, ~40 nm, expected particle diameter.

    Techniques Used: Infection, Derivative Assay, Control, Cell Culture, Virus, Quantitative RT-PCR

    ( A ) J4FUT2-KI HIEs were pretreated with 30 μM TAK-779 and then infected with GII.3 (TCH-04-577/P21, 2.9 × 10 5 GE per well; TCH-25-396/P12, 2.2 × 10 5 GE per well), GII.17 (1295-44/P13, 2.8 × 10 5 GE per well; TCH-25-256/P171, 3.1 × 10 5 GE per well), G1.1 (BCM-723–100595/P1, 3.7 × 10 5 GEs per well; 51899/P1, 6.9 × 10 5 GE per well). ( B ) J4FUT2-KI HIEs were pretreated with 30 μM TAK-779 and then infected with GII.4 Sydney (BCM 16-2/P31, 3.0 × 10 5 GE per well; TCH-24-163/P16, 2.2 × 10 5 GE per well), GII.4 Den Haag (TCH-23-323/P16, 5.6 × 10 6 GE per well), GII.4 Hunter (TCH-05-797/P4, 3.35 × 10 5 GE per well) in the absence or presence of 30 μM TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of 30 μM TAK-779. Virus replication at 1 and 48 hours postinfection was evaluated by RT-qPCR. Mean data compiled from two independent experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors Error bars denote SD. Significance was determined using paired t test comparing 48-hour replication, 0 to 30 μM TAK-779 ( P value, *, 0.01; **, 0.01; ***, 0.001; and ****, < 0.0001). hr, hours.
    Figure Legend Snippet: ( A ) J4FUT2-KI HIEs were pretreated with 30 μM TAK-779 and then infected with GII.3 (TCH-04-577/P21, 2.9 × 10 5 GE per well; TCH-25-396/P12, 2.2 × 10 5 GE per well), GII.17 (1295-44/P13, 2.8 × 10 5 GE per well; TCH-25-256/P171, 3.1 × 10 5 GE per well), G1.1 (BCM-723–100595/P1, 3.7 × 10 5 GEs per well; 51899/P1, 6.9 × 10 5 GE per well). ( B ) J4FUT2-KI HIEs were pretreated with 30 μM TAK-779 and then infected with GII.4 Sydney (BCM 16-2/P31, 3.0 × 10 5 GE per well; TCH-24-163/P16, 2.2 × 10 5 GE per well), GII.4 Den Haag (TCH-23-323/P16, 5.6 × 10 6 GE per well), GII.4 Hunter (TCH-05-797/P4, 3.35 × 10 5 GE per well) in the absence or presence of 30 μM TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of 30 μM TAK-779. Virus replication at 1 and 48 hours postinfection was evaluated by RT-qPCR. Mean data compiled from two independent experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors Error bars denote SD. Significance was determined using paired t test comparing 48-hour replication, 0 to 30 μM TAK-779 ( P value, *, 0.01; **, 0.01; ***, 0.001; and ****, < 0.0001). hr, hours.

    Techniques Used: Infection, Cell Culture, Virus, Quantitative RT-PCR



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    MedChemExpress tak 1 inhib hs 276
    A. Quantifications of ASC-GFP specks in HEK293 ASC-GFP/NLRP1 reporter cells exposed to Dinophysis toxin (100nM) or not for 6 h in presence or absence of various MAPK inhibitors (10µM). TAK1 inhibitor; <t>HS-276,</t> ZAKα inhibitor; PLX4720, TAOK inhibitor; CP-43, MLKL inhibitor; necro sulfonamide, ASK1 inhibitor; GS-444217, DLK/LZK inhibitor; DN-1289, RIPK3 inhibitor; GSK-872. ASC-GFP (green) pictures were directly taken in dish after adding Hoechst (nuclei staining). Images shown are from one experiment and are representative of three independent experiments; scale bars, 10 µm. ASC complex percentage was performed by determining the ratios of cells positive for ASC speckles on the total nuclei (Hoechst). At least ten fields from each experiment were analyzed. Values are expressed as mean ± SEM. ***P ≤ 0.0001, one-way ANOVA. B. Determination of the IL-1β release in WT, P38α/β/δ TKO, TAK1 KO and NLRP1 KO NTERT keratinocytes exposed or not to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM), Anisomycin (1µM) and VbP (10µM) for 10 hours. ***P ≤ 0.0001, one-way ANOVA. Values are expressed as mean ± SEM. Graphs show one experiment performed in triplicates at least three times. C. Immunoblotting of P38, TAK1, cleaved GSDMD and IL-1β and of the subsequent IL-1β release in WT, P38α/β/δ TKO, TAK1 KO, P38α/β/δ TKO/TAK1 KO and NLRP1 KO NTERT keratinocytes exposed or not to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM), Anisomycin (1µM) and VbP (10µM) for 10 hours. ***P ≤ 0.0001, one-way ANOVA. Values are expressed as mean ± SEM. Graphs show one experiment performed in triplicates at least three times. D. Phosphotag blotting of phosphorylated P38, TAK1 and NLRP1-DR-SNAP in WT, P38α/β/δ TKO, TAK1 KO or P38α/β/δ TKO/TAK1 KO NTERT keratinocytes exposed to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM) for 1 hour. Tubulin-α was used as internal protein loading controls. Immunoblots show lysates from one experiment performed at least three times. E. Phosphotag immunoblotting of phosphorylated recombinant NLRP1 full-length protein or immunoprecipitated GFP-tagged NLRP1-DR incubated with recombinant TAK1-TAB1 fusion or P38α kinases for 60 minutes in presence/absence of lambda phosphatase. Immunoblots show proteins from one experiment performed at least three times. F. Fluorescence microscopy quantifications of ASC-GFP specks in WT, P38α/β/δ TKO or in P38α/β/δ TKO/TAK1 KO HEK293 ASC-GFP/NLRP1 reporter cells after 6 h of exposure to Dinophysis toxin (100nM) or Val-boro-Pro (VbP, 10µM). ASC complex percentage was performed by determining the ratios of cells positive for ASC speckles (green, GFP) on the total nuclei (Hoechst). At least ten fields from three independent experiments were analyzed. Values are expressed as mean ± SEM. ***P ≤ 0.0001, two-way ANOVA with multiple comparisons. Graphs show one experiment performed in triplicate at least three times.
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    ( A to C ) HIEs were pretreated for 3 hours with the indicated concentrations of TAK-779 and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the absence or presence of the same concentrations of TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from two independent experiments (J2 and J8FUT2-KI HIEs) and three independent experiments (J4FUT2-KI HIEs) with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Significance was determined using two-way ANOVA with Tukey’s post hoc test comparing TAK-779 concentrations and individual experimental results at 48 hours postinfection. P values are shown for individual TAK-779 concentrations versus no TAK-779 treatment control (* P value <0.05, ** P < 0.01, *** P < 0.001, and ****, 0.0001). Significant differences between experiments were observed for the studies conducted in J4FUT2-KI and J8FUT2-KI HIEs ( P < 0.0001 for each).

    Journal: Science Advances

    Article Title: Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids

    doi: 10.1126/sciadv.aeb0455

    Figure Lengend Snippet: ( A to C ) HIEs were pretreated for 3 hours with the indicated concentrations of TAK-779 and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the absence or presence of the same concentrations of TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from two independent experiments (J2 and J8FUT2-KI HIEs) and three independent experiments (J4FUT2-KI HIEs) with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Significance was determined using two-way ANOVA with Tukey’s post hoc test comparing TAK-779 concentrations and individual experimental results at 48 hours postinfection. P values are shown for individual TAK-779 concentrations versus no TAK-779 treatment control (* P value <0.05, ** P < 0.01, *** P < 0.001, and ****, 0.0001). Significant differences between experiments were observed for the studies conducted in J4FUT2-KI and J8FUT2-KI HIEs ( P < 0.0001 for each).

    Article Snippet: TAK-779 was purchased from Medchem Express (catalog no. HY-13406) and reconstituted in water to make 5 mM stocks and frozen in single-use aliquots to avoid repeated freeze-thaw cycles.

    Techniques: Infection, Cell Culture, Quantitative RT-PCR, Control

    Different conditions, pre- versus post-treatment (indicated) with pretreatment alone compared to post-treatment alone and 24 hours pretreatment, were compared. J4FUT2-KI HIEs were treated as indicated with 30 μM TAK-779. HIEs were then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the indicated conditions. After washing, the cells were cultured for 48 hours at 37 ° C in the presence of 30 μM TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from three independent experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Numbers above lines represent log 10 increase between no treatment versus indicated treatments. Significance was determined using two-way ANOVA with post hoc Tukey’s test comparing 48-hour replication, no TAK-779 to each condition of TAK-779 ( P value,**, < 0.001 and ****, 0.0001). hr, hours.

    Journal: Science Advances

    Article Title: Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids

    doi: 10.1126/sciadv.aeb0455

    Figure Lengend Snippet: Different conditions, pre- versus post-treatment (indicated) with pretreatment alone compared to post-treatment alone and 24 hours pretreatment, were compared. J4FUT2-KI HIEs were treated as indicated with 30 μM TAK-779. HIEs were then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the indicated conditions. After washing, the cells were cultured for 48 hours at 37 ° C in the presence of 30 μM TAK-779. Viral GEs at 1 and 48 hours postinfection were quantified by RT-qPCR. Mean data compiled from three independent experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors. Numbers above lines represent log 10 increase between no treatment versus indicated treatments. Significance was determined using two-way ANOVA with post hoc Tukey’s test comparing 48-hour replication, no TAK-779 to each condition of TAK-779 ( P value,**, < 0.001 and ****, 0.0001). hr, hours.

    Article Snippet: TAK-779 was purchased from Medchem Express (catalog no. HY-13406) and reconstituted in water to make 5 mM stocks and frozen in single-use aliquots to avoid repeated freeze-thaw cycles.

    Techniques: Infection, Cell Culture, Quantitative RT-PCR

    ( A ) J4FUT2-KI HIEs were pretreated for 3 hours and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the presence of the indicated concentrations of TAK-779. After washing, the cells were cultured for the indicated times at 37°C in the presence of the indicated concentrations of TAK-779. Virus replication was evaluated by RT-qPCR at the indicated time points. ( B and C ) After 24 and 48 hours postinfection incubation, monolayers were fixed with methanol. GII.3-positive cells (green) were detected using guinea pig anti-HuNoV VLP Ab (green) and nuclei (blue) were detected by DAPI. (B) Representative images from each condition at 48 hours postinfection are shown. Scale bars denote 14 μm. (C) Numbers of HuNoV infected cells (Foci) per well from 24 and 48 hours postinfection condition were counted. Mean data compiled from two experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors using one-way ANOVA with post hoc Newman Keuls test comparing GEs per well for TAK-779 concentrations at 48- and 72-hour time points, ( P value, ****, 0.0001).

    Journal: Science Advances

    Article Title: Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids

    doi: 10.1126/sciadv.aeb0455

    Figure Lengend Snippet: ( A ) J4FUT2-KI HIEs were pretreated for 3 hours and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the presence of the indicated concentrations of TAK-779. After washing, the cells were cultured for the indicated times at 37°C in the presence of the indicated concentrations of TAK-779. Virus replication was evaluated by RT-qPCR at the indicated time points. ( B and C ) After 24 and 48 hours postinfection incubation, monolayers were fixed with methanol. GII.3-positive cells (green) were detected using guinea pig anti-HuNoV VLP Ab (green) and nuclei (blue) were detected by DAPI. (B) Representative images from each condition at 48 hours postinfection are shown. Scale bars denote 14 μm. (C) Numbers of HuNoV infected cells (Foci) per well from 24 and 48 hours postinfection condition were counted. Mean data compiled from two experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors using one-way ANOVA with post hoc Newman Keuls test comparing GEs per well for TAK-779 concentrations at 48- and 72-hour time points, ( P value, ****, 0.0001).

    Article Snippet: TAK-779 was purchased from Medchem Express (catalog no. HY-13406) and reconstituted in water to make 5 mM stocks and frozen in single-use aliquots to avoid repeated freeze-thaw cycles.

    Techniques: Infection, Cell Culture, Virus, Quantitative RT-PCR, Incubation

    ( A ) J4FUT2-KI and ( B ) J8FUT2-KI HIEs were pretreated for 3 hours and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) with or without 30 μM TAK-779, labeled as P1. Following infection, both cells and supernatant were collected at 96 hours postinfection, and a portion was used to quantify viral RNA by RT-qPCR. The virus was subsequently used to infect a fresh batch of HIEs for the next passage, continuing for up to five passages. dpi, days postinfection; hpi, hours postinfection.

    Journal: Science Advances

    Article Title: Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids

    doi: 10.1126/sciadv.aeb0455

    Figure Lengend Snippet: ( A ) J4FUT2-KI and ( B ) J8FUT2-KI HIEs were pretreated for 3 hours and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) with or without 30 μM TAK-779, labeled as P1. Following infection, both cells and supernatant were collected at 96 hours postinfection, and a portion was used to quantify viral RNA by RT-qPCR. The virus was subsequently used to infect a fresh batch of HIEs for the next passage, continuing for up to five passages. dpi, days postinfection; hpi, hours postinfection.

    Article Snippet: TAK-779 was purchased from Medchem Express (catalog no. HY-13406) and reconstituted in water to make 5 mM stocks and frozen in single-use aliquots to avoid repeated freeze-thaw cycles.

    Techniques: Infection, Labeling, Quantitative RT-PCR, Virus

    ( A ) J4FUT2-KI and J8FUT2-KI HIEs were pretreated and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the presence of 30 μM TAK-779 and indicated as P1. Postinfection, the cells and supernatant were harvested at 96 hours postinfection; after processing the supernatant, an aliquot was used to quantify the viral RNA by RT-qPCR. The virus was then used to infect a new batch of HIEs for the next passage, continuing up to 10 passages. ( B ) Immunofluorescence staining of HuNoV-infected J4FUT2-KI monolayers at P5 and P2. At 48 hours postinfection, cells were fixed with methanol and stained with guinea pig anti-HuNoV VLP antibody (green), rabbit anti-NTPase (red), and mouse anti-VPg (white). Nuclei were counterstained with DAPI (blue). Representative images are shown at high (10-μm scale bar) and low (20-μm scale bar) magnification. dpi, days postinfection; hpi, hours postinfection.

    Journal: Science Advances

    Article Title: Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids

    doi: 10.1126/sciadv.aeb0455

    Figure Lengend Snippet: ( A ) J4FUT2-KI and J8FUT2-KI HIEs were pretreated and then infected with GII.3 HuNoV (2.9 × 10 5 GEs per well) in the presence of 30 μM TAK-779 and indicated as P1. Postinfection, the cells and supernatant were harvested at 96 hours postinfection; after processing the supernatant, an aliquot was used to quantify the viral RNA by RT-qPCR. The virus was then used to infect a new batch of HIEs for the next passage, continuing up to 10 passages. ( B ) Immunofluorescence staining of HuNoV-infected J4FUT2-KI monolayers at P5 and P2. At 48 hours postinfection, cells were fixed with methanol and stained with guinea pig anti-HuNoV VLP antibody (green), rabbit anti-NTPase (red), and mouse anti-VPg (white). Nuclei were counterstained with DAPI (blue). Representative images are shown at high (10-μm scale bar) and low (20-μm scale bar) magnification. dpi, days postinfection; hpi, hours postinfection.

    Article Snippet: TAK-779 was purchased from Medchem Express (catalog no. HY-13406) and reconstituted in water to make 5 mM stocks and frozen in single-use aliquots to avoid repeated freeze-thaw cycles.

    Techniques: Infection, Quantitative RT-PCR, Virus, Immunofluorescence, Staining

    ( A ) J4FUT2-KI HIEs were pretreated for 3 hours with 30 μM TAK-779 and infected with GII.3 HuNoV derived from stool [2.9 × 10 5 genome equivalents (GEs) per well] or from stock 1 (2.58 × 10 5 GEs per well), stock 2 (2.93 × 10 5 GEs per well), stock 3 (1.02 × 10 5 GEs per well), or stock 4 (2.50 × 10 5 GEs per well). Infections were performed in the presence of 30 μM TAK-779 (left) or vehicle control (right). After washing, the cells were cultured for the indicated times at 37°C in the presence (left) or absence (right) of TAK-779. Virus replication was evaluated by RT-qPCR at the indicated time points. Viral replication was assessed by RT-qPCR. The characterization of each stock was performed once with three replicates. Error bars denote SD. ( B ) Representative images of stock-infected HIEs fixed with methanol at 48 hours postinfection. GII.3-positive cells were detected using guinea pig anti-HuNoV VLP Ab (green) and nuclei (blue) were detected by DAPI. Scale bars denote 14 μm. ( C ) Electron micrograph of HuNoV particles from the supernatant of HIEs 24 hours postinfection infected with stock virus. Scale bar, 50 nm; red line, ~40 nm, expected particle diameter.

    Journal: Science Advances

    Article Title: Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids

    doi: 10.1126/sciadv.aeb0455

    Figure Lengend Snippet: ( A ) J4FUT2-KI HIEs were pretreated for 3 hours with 30 μM TAK-779 and infected with GII.3 HuNoV derived from stool [2.9 × 10 5 genome equivalents (GEs) per well] or from stock 1 (2.58 × 10 5 GEs per well), stock 2 (2.93 × 10 5 GEs per well), stock 3 (1.02 × 10 5 GEs per well), or stock 4 (2.50 × 10 5 GEs per well). Infections were performed in the presence of 30 μM TAK-779 (left) or vehicle control (right). After washing, the cells were cultured for the indicated times at 37°C in the presence (left) or absence (right) of TAK-779. Virus replication was evaluated by RT-qPCR at the indicated time points. Viral replication was assessed by RT-qPCR. The characterization of each stock was performed once with three replicates. Error bars denote SD. ( B ) Representative images of stock-infected HIEs fixed with methanol at 48 hours postinfection. GII.3-positive cells were detected using guinea pig anti-HuNoV VLP Ab (green) and nuclei (blue) were detected by DAPI. Scale bars denote 14 μm. ( C ) Electron micrograph of HuNoV particles from the supernatant of HIEs 24 hours postinfection infected with stock virus. Scale bar, 50 nm; red line, ~40 nm, expected particle diameter.

    Article Snippet: TAK-779 was purchased from Medchem Express (catalog no. HY-13406) and reconstituted in water to make 5 mM stocks and frozen in single-use aliquots to avoid repeated freeze-thaw cycles.

    Techniques: Infection, Derivative Assay, Control, Cell Culture, Virus, Quantitative RT-PCR

    ( A ) J4FUT2-KI HIEs were pretreated with 30 μM TAK-779 and then infected with GII.3 (TCH-04-577/P21, 2.9 × 10 5 GE per well; TCH-25-396/P12, 2.2 × 10 5 GE per well), GII.17 (1295-44/P13, 2.8 × 10 5 GE per well; TCH-25-256/P171, 3.1 × 10 5 GE per well), G1.1 (BCM-723–100595/P1, 3.7 × 10 5 GEs per well; 51899/P1, 6.9 × 10 5 GE per well). ( B ) J4FUT2-KI HIEs were pretreated with 30 μM TAK-779 and then infected with GII.4 Sydney (BCM 16-2/P31, 3.0 × 10 5 GE per well; TCH-24-163/P16, 2.2 × 10 5 GE per well), GII.4 Den Haag (TCH-23-323/P16, 5.6 × 10 6 GE per well), GII.4 Hunter (TCH-05-797/P4, 3.35 × 10 5 GE per well) in the absence or presence of 30 μM TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of 30 μM TAK-779. Virus replication at 1 and 48 hours postinfection was evaluated by RT-qPCR. Mean data compiled from two independent experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors Error bars denote SD. Significance was determined using paired t test comparing 48-hour replication, 0 to 30 μM TAK-779 ( P value, *, 0.01; **, 0.01; ***, 0.001; and ****, < 0.0001). hr, hours.

    Journal: Science Advances

    Article Title: Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids

    doi: 10.1126/sciadv.aeb0455

    Figure Lengend Snippet: ( A ) J4FUT2-KI HIEs were pretreated with 30 μM TAK-779 and then infected with GII.3 (TCH-04-577/P21, 2.9 × 10 5 GE per well; TCH-25-396/P12, 2.2 × 10 5 GE per well), GII.17 (1295-44/P13, 2.8 × 10 5 GE per well; TCH-25-256/P171, 3.1 × 10 5 GE per well), G1.1 (BCM-723–100595/P1, 3.7 × 10 5 GEs per well; 51899/P1, 6.9 × 10 5 GE per well). ( B ) J4FUT2-KI HIEs were pretreated with 30 μM TAK-779 and then infected with GII.4 Sydney (BCM 16-2/P31, 3.0 × 10 5 GE per well; TCH-24-163/P16, 2.2 × 10 5 GE per well), GII.4 Den Haag (TCH-23-323/P16, 5.6 × 10 6 GE per well), GII.4 Hunter (TCH-05-797/P4, 3.35 × 10 5 GE per well) in the absence or presence of 30 μM TAK-779. After washing, the cells were cultured for 48 hours at 37°C in the absence or presence of 30 μM TAK-779. Virus replication at 1 and 48 hours postinfection was evaluated by RT-qPCR. Mean data compiled from two independent experiments with three wells per experiment are shown; error bars show SD. Experiments are denoted with different symbol colors Error bars denote SD. Significance was determined using paired t test comparing 48-hour replication, 0 to 30 μM TAK-779 ( P value, *, 0.01; **, 0.01; ***, 0.001; and ****, < 0.0001). hr, hours.

    Article Snippet: TAK-779 was purchased from Medchem Express (catalog no. HY-13406) and reconstituted in water to make 5 mM stocks and frozen in single-use aliquots to avoid repeated freeze-thaw cycles.

    Techniques: Infection, Cell Culture, Virus, Quantitative RT-PCR

    ( A ) The peptide library consists of 214,158 different 30-residue partially overlapping fragments covering 5672 cytosolic human proteins. Parts of the figure were created in BioRender. R. Hartmann-Petersen (2025), https://BioRender.com/23ydezu . ( B ) Schematic illustration of the expression and screening system. The plasmid containing the GFP-fused peptide library also includes an internal ribosomal entry site (IRES), mCherry, and a site for Bxb1 recombination into a landing pad in HEK293T cells. Expression from the landing pad is regulated by the Tet-on promoter (bent arrow), which, in nonrecombined cells, drives the expression of BFP, inducible caspase 9 (iCasp9), and a blasticidin resistance gene (Blast R ) separated with a parechovirus 2A–like translational stop-start sequence (2A). Upon correct integration, the GFP fragments and mCherry are expressed from the same mRNA. Using fluorescence-activated cell sorting (FACS), cells are sorted into four equally populated bins, and the fragments in each bin can be identified by sequencing. Parts of the figure were created in BioRender. R. Hartmann-Petersen (2025), https://BioRender.com/qw20nnx and adapted from ( , , ). Representative distributions of GFP:mCherry ratios in cells expressing the library and either untreated (control) or ( C ) treated for 8 hours with cycloheximide (CHX; 10 μg/ml) ( n = 900,000), ( D ) for 16 hours with 15 μM bortezomib (BZ) ( n = 720,000), or ( E ) 16 hours with 1 μM MLN7243 ( n = 347,077; untreated, n = 340,000). ( F ) A representative flow cytometry (FC) profile for cells expressing the library ( n = 495,000). Bin thresholds used to sort the library into four (bin 1 to bin 4) equally populated bins (25% in each bin) are shown as black horizontal bars.

    Journal: Science Advances

    Article Title: A near-complete map of human cytosolic degrons and their relevance for disease

    doi: 10.1126/sciadv.adz3483

    Figure Lengend Snippet: ( A ) The peptide library consists of 214,158 different 30-residue partially overlapping fragments covering 5672 cytosolic human proteins. Parts of the figure were created in BioRender. R. Hartmann-Petersen (2025), https://BioRender.com/23ydezu . ( B ) Schematic illustration of the expression and screening system. The plasmid containing the GFP-fused peptide library also includes an internal ribosomal entry site (IRES), mCherry, and a site for Bxb1 recombination into a landing pad in HEK293T cells. Expression from the landing pad is regulated by the Tet-on promoter (bent arrow), which, in nonrecombined cells, drives the expression of BFP, inducible caspase 9 (iCasp9), and a blasticidin resistance gene (Blast R ) separated with a parechovirus 2A–like translational stop-start sequence (2A). Upon correct integration, the GFP fragments and mCherry are expressed from the same mRNA. Using fluorescence-activated cell sorting (FACS), cells are sorted into four equally populated bins, and the fragments in each bin can be identified by sequencing. Parts of the figure were created in BioRender. R. Hartmann-Petersen (2025), https://BioRender.com/qw20nnx and adapted from ( , , ). Representative distributions of GFP:mCherry ratios in cells expressing the library and either untreated (control) or ( C ) treated for 8 hours with cycloheximide (CHX; 10 μg/ml) ( n = 900,000), ( D ) for 16 hours with 15 μM bortezomib (BZ) ( n = 720,000), or ( E ) 16 hours with 1 μM MLN7243 ( n = 347,077; untreated, n = 340,000). ( F ) A representative flow cytometry (FC) profile for cells expressing the library ( n = 495,000). Bin thresholds used to sort the library into four (bin 1 to bin 4) equally populated bins (25% in each bin) are shown as black horizontal bars.

    Article Snippet: Last, the sublibrary was treated with 1 μM MLN7243 (MedChemExpress) and 1.5 μM MLN4924 (MedChemExpress) for 16 hours.

    Techniques: Residue, Expressing, Plasmid Preparation, Sequencing, Fluorescence, FACS, Control, Flow Cytometry

    A. Quantifications of ASC-GFP specks in HEK293 ASC-GFP/NLRP1 reporter cells exposed to Dinophysis toxin (100nM) or not for 6 h in presence or absence of various MAPK inhibitors (10µM). TAK1 inhibitor; HS-276, ZAKα inhibitor; PLX4720, TAOK inhibitor; CP-43, MLKL inhibitor; necro sulfonamide, ASK1 inhibitor; GS-444217, DLK/LZK inhibitor; DN-1289, RIPK3 inhibitor; GSK-872. ASC-GFP (green) pictures were directly taken in dish after adding Hoechst (nuclei staining). Images shown are from one experiment and are representative of three independent experiments; scale bars, 10 µm. ASC complex percentage was performed by determining the ratios of cells positive for ASC speckles on the total nuclei (Hoechst). At least ten fields from each experiment were analyzed. Values are expressed as mean ± SEM. ***P ≤ 0.0001, one-way ANOVA. B. Determination of the IL-1β release in WT, P38α/β/δ TKO, TAK1 KO and NLRP1 KO NTERT keratinocytes exposed or not to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM), Anisomycin (1µM) and VbP (10µM) for 10 hours. ***P ≤ 0.0001, one-way ANOVA. Values are expressed as mean ± SEM. Graphs show one experiment performed in triplicates at least three times. C. Immunoblotting of P38, TAK1, cleaved GSDMD and IL-1β and of the subsequent IL-1β release in WT, P38α/β/δ TKO, TAK1 KO, P38α/β/δ TKO/TAK1 KO and NLRP1 KO NTERT keratinocytes exposed or not to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM), Anisomycin (1µM) and VbP (10µM) for 10 hours. ***P ≤ 0.0001, one-way ANOVA. Values are expressed as mean ± SEM. Graphs show one experiment performed in triplicates at least three times. D. Phosphotag blotting of phosphorylated P38, TAK1 and NLRP1-DR-SNAP in WT, P38α/β/δ TKO, TAK1 KO or P38α/β/δ TKO/TAK1 KO NTERT keratinocytes exposed to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM) for 1 hour. Tubulin-α was used as internal protein loading controls. Immunoblots show lysates from one experiment performed at least three times. E. Phosphotag immunoblotting of phosphorylated recombinant NLRP1 full-length protein or immunoprecipitated GFP-tagged NLRP1-DR incubated with recombinant TAK1-TAB1 fusion or P38α kinases for 60 minutes in presence/absence of lambda phosphatase. Immunoblots show proteins from one experiment performed at least three times. F. Fluorescence microscopy quantifications of ASC-GFP specks in WT, P38α/β/δ TKO or in P38α/β/δ TKO/TAK1 KO HEK293 ASC-GFP/NLRP1 reporter cells after 6 h of exposure to Dinophysis toxin (100nM) or Val-boro-Pro (VbP, 10µM). ASC complex percentage was performed by determining the ratios of cells positive for ASC speckles (green, GFP) on the total nuclei (Hoechst). At least ten fields from three independent experiments were analyzed. Values are expressed as mean ± SEM. ***P ≤ 0.0001, two-way ANOVA with multiple comparisons. Graphs show one experiment performed in triplicate at least three times.

    Journal: bioRxiv

    Article Title: A TAK1-Driven NLRP1 Inflammasome Pathway Revealed by Phosphatase-Targeting Environmental Toxins

    doi: 10.64898/2026.01.23.701233

    Figure Lengend Snippet: A. Quantifications of ASC-GFP specks in HEK293 ASC-GFP/NLRP1 reporter cells exposed to Dinophysis toxin (100nM) or not for 6 h in presence or absence of various MAPK inhibitors (10µM). TAK1 inhibitor; HS-276, ZAKα inhibitor; PLX4720, TAOK inhibitor; CP-43, MLKL inhibitor; necro sulfonamide, ASK1 inhibitor; GS-444217, DLK/LZK inhibitor; DN-1289, RIPK3 inhibitor; GSK-872. ASC-GFP (green) pictures were directly taken in dish after adding Hoechst (nuclei staining). Images shown are from one experiment and are representative of three independent experiments; scale bars, 10 µm. ASC complex percentage was performed by determining the ratios of cells positive for ASC speckles on the total nuclei (Hoechst). At least ten fields from each experiment were analyzed. Values are expressed as mean ± SEM. ***P ≤ 0.0001, one-way ANOVA. B. Determination of the IL-1β release in WT, P38α/β/δ TKO, TAK1 KO and NLRP1 KO NTERT keratinocytes exposed or not to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM), Anisomycin (1µM) and VbP (10µM) for 10 hours. ***P ≤ 0.0001, one-way ANOVA. Values are expressed as mean ± SEM. Graphs show one experiment performed in triplicates at least three times. C. Immunoblotting of P38, TAK1, cleaved GSDMD and IL-1β and of the subsequent IL-1β release in WT, P38α/β/δ TKO, TAK1 KO, P38α/β/δ TKO/TAK1 KO and NLRP1 KO NTERT keratinocytes exposed or not to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM), Anisomycin (1µM) and VbP (10µM) for 10 hours. ***P ≤ 0.0001, one-way ANOVA. Values are expressed as mean ± SEM. Graphs show one experiment performed in triplicates at least three times. D. Phosphotag blotting of phosphorylated P38, TAK1 and NLRP1-DR-SNAP in WT, P38α/β/δ TKO, TAK1 KO or P38α/β/δ TKO/TAK1 KO NTERT keratinocytes exposed to Dinophysis toxin (Dino. toxin, 100nM), Okadaic acid (Ok. Acid, 250nM), Cantharidin (Canth. 5µM) for 1 hour. Tubulin-α was used as internal protein loading controls. Immunoblots show lysates from one experiment performed at least three times. E. Phosphotag immunoblotting of phosphorylated recombinant NLRP1 full-length protein or immunoprecipitated GFP-tagged NLRP1-DR incubated with recombinant TAK1-TAB1 fusion or P38α kinases for 60 minutes in presence/absence of lambda phosphatase. Immunoblots show proteins from one experiment performed at least three times. F. Fluorescence microscopy quantifications of ASC-GFP specks in WT, P38α/β/δ TKO or in P38α/β/δ TKO/TAK1 KO HEK293 ASC-GFP/NLRP1 reporter cells after 6 h of exposure to Dinophysis toxin (100nM) or Val-boro-Pro (VbP, 10µM). ASC complex percentage was performed by determining the ratios of cells positive for ASC speckles (green, GFP) on the total nuclei (Hoechst). At least ten fields from three independent experiments were analyzed. Values are expressed as mean ± SEM. ***P ≤ 0.0001, two-way ANOVA with multiple comparisons. Graphs show one experiment performed in triplicate at least three times.

    Article Snippet: If not otherwise indicated, stimuli and inhibitors were used at the following concentrations: PLX-4720 (10 μM, MedChem Express, HY-51424); SB203580 (10 μM, MedChem Express, HY-10256); Doramapimod (10 μM, MedChem Express, HY-10320); BX795 (1 μM, Invivogen, Tlrl-bx7); Abemaciclib (25 nM, MedChem Express, HY-16297A); Ribociclib (40 nM, MedChem Express, HY-15777); Palbociclib (25 nM, MedChem Express, HY-50767); Fadraciclib (25 nM, MedChem Express, HY-101212); Roniciclib (25 nM, MedChem Express, HY-13914); SCH772984 (1 μM, MedChem Express, HY-50846); SP600125 (10 μM, MedChem Express, HY-12041); TAK-1 inhib HS-276 (10-20 μM, MedChem Express, HY-147141); TAO Kinase inhibitor 1 (5 μM, MedChem Express, HY-112136); Selonsertib (10 μM, MedChem Express, HY-18938); 5z-7-Oxozeaenol (5 μM, MedChem Express, HY-12686); GW806742X (1 μM, MedChem Express, HY-112292A); DN1289 (1 μM, MedChem Express, HY-152142); Gossypetin (40 μM, MedChem Express, HY-119917); BSJ-04-122 (10 μM, MedChem Express, HY-152185); GSK-872 (10 μM, MedChem Express, HY-101872); Bortezomib (1 μM, Selleck, SE-S1013-5MG); MLN9424 (1 μM, Tocris Bioscience, 6499); DT-061 (1-20 μM; MedChem Express, HY-112929); Dinophysistoxin (250 nM, Bertin Technologies); Okadaic acid (100-600 nM, Bertin Technologies, WP-10011490); Cantharidin (5-25 μM, Sigma-Aldrich, C7632-25MG); Calyculin-A (1 μM, Bertin Technologies, WP-19246); LB-100 (50 μM, Bertin Technologies, 29105); Raphin-1 (50 μM, Tocris Bioscience, 6760/10); Sephin-1 (100 μM, Tocris Bioscience, 5553/10); Anisomycin (1-5 μM, Selleck, SE-S7409-10MG); Valboro-Pro/ Talabostat (10 μM, Selleck, SE-S8455-5MG); Phosphatase Inhibitor Library (117 items) (10 μM, MedChem Express, HY-L081).

    Techniques: Staining, Western Blot, Recombinant, Immunoprecipitation, Incubation, Fluorescence, Microscopy

    A-C . Hematoxylin (H) & Eosin (E) or ASC immunobiological staining showing P38 and TAK1-dependent histological/inflammasome changes caused by Dinophysis toxin (250 nM), Okadaic acid (600 nM) and Cantharidin (10µM) exposure for 24 hours. When specified, pan P38 kinase inhibitor Doramapimod (10µM) and TAK1 inhibitor HS-276 (20µM) were used. Associated quantification of IL-1β release, the dermal–epidermal layer detachment and the percentage of ASC specks in the Human skin explants. P values indicated in figure, one-way ANOVA. Images are representative of two (A) and three (B) biological replicates. Scale bar = 50 µm.

    Journal: bioRxiv

    Article Title: A TAK1-Driven NLRP1 Inflammasome Pathway Revealed by Phosphatase-Targeting Environmental Toxins

    doi: 10.64898/2026.01.23.701233

    Figure Lengend Snippet: A-C . Hematoxylin (H) & Eosin (E) or ASC immunobiological staining showing P38 and TAK1-dependent histological/inflammasome changes caused by Dinophysis toxin (250 nM), Okadaic acid (600 nM) and Cantharidin (10µM) exposure for 24 hours. When specified, pan P38 kinase inhibitor Doramapimod (10µM) and TAK1 inhibitor HS-276 (20µM) were used. Associated quantification of IL-1β release, the dermal–epidermal layer detachment and the percentage of ASC specks in the Human skin explants. P values indicated in figure, one-way ANOVA. Images are representative of two (A) and three (B) biological replicates. Scale bar = 50 µm.

    Article Snippet: If not otherwise indicated, stimuli and inhibitors were used at the following concentrations: PLX-4720 (10 μM, MedChem Express, HY-51424); SB203580 (10 μM, MedChem Express, HY-10256); Doramapimod (10 μM, MedChem Express, HY-10320); BX795 (1 μM, Invivogen, Tlrl-bx7); Abemaciclib (25 nM, MedChem Express, HY-16297A); Ribociclib (40 nM, MedChem Express, HY-15777); Palbociclib (25 nM, MedChem Express, HY-50767); Fadraciclib (25 nM, MedChem Express, HY-101212); Roniciclib (25 nM, MedChem Express, HY-13914); SCH772984 (1 μM, MedChem Express, HY-50846); SP600125 (10 μM, MedChem Express, HY-12041); TAK-1 inhib HS-276 (10-20 μM, MedChem Express, HY-147141); TAO Kinase inhibitor 1 (5 μM, MedChem Express, HY-112136); Selonsertib (10 μM, MedChem Express, HY-18938); 5z-7-Oxozeaenol (5 μM, MedChem Express, HY-12686); GW806742X (1 μM, MedChem Express, HY-112292A); DN1289 (1 μM, MedChem Express, HY-152142); Gossypetin (40 μM, MedChem Express, HY-119917); BSJ-04-122 (10 μM, MedChem Express, HY-152185); GSK-872 (10 μM, MedChem Express, HY-101872); Bortezomib (1 μM, Selleck, SE-S1013-5MG); MLN9424 (1 μM, Tocris Bioscience, 6499); DT-061 (1-20 μM; MedChem Express, HY-112929); Dinophysistoxin (250 nM, Bertin Technologies); Okadaic acid (100-600 nM, Bertin Technologies, WP-10011490); Cantharidin (5-25 μM, Sigma-Aldrich, C7632-25MG); Calyculin-A (1 μM, Bertin Technologies, WP-19246); LB-100 (50 μM, Bertin Technologies, 29105); Raphin-1 (50 μM, Tocris Bioscience, 6760/10); Sephin-1 (100 μM, Tocris Bioscience, 5553/10); Anisomycin (1-5 μM, Selleck, SE-S7409-10MG); Valboro-Pro/ Talabostat (10 μM, Selleck, SE-S8455-5MG); Phosphatase Inhibitor Library (117 items) (10 μM, MedChem Express, HY-L081).

    Techniques: Staining